Plodia interpunctella (Lepidoptera: Pyralidae): Intoxication with essential oils isolated from Lippia turbinata (Griseb.) and analysis of neuropeptides and neuropeptide receptors,putative targets for pest control

The Indian meal moth Plodia interpunctella is a pest of stored products worldwide. Plant-derived essential oils with insecticidal activity could be safe products to control this species. The scarce information about the mode of action of most plant-derived products limits their use for the control of insect pests. Here, we demonstrate that an essential oil distilled from Lippia turbinata (“poleo”) has insecticidal activity on P. interpunctella larvae. Furthermore, we performed a comprehensive characterization of P. interpunctella neuroendocrine system, in comparison with other lepidopteran species.     

Evaluation of the composition and fumigant toxicity against Plodia interpunctella of essential oils from Ajania potaninii and Ajania fruticulosa

Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae), is a worldwide omnivorous pest. It is the primary insect pest in many economically important stored crops. The insecticidal activity of essential oils (EOs) extracted from Ajania potaninii and Ajania fruticulosa were evaluated against Plodia interpunctella. EOs obtained by hydro-distillation were analyzed by GC–MS. Fumigant toxicity testing indicated that both EOs and their main components were toxic to P. interpunctella adults. 1,8-Cineole exhibited the strongest activity, having an LC₅₀ of 0.86 mg/L air and being twice as active as camphor. Myrtenol was also strongly toxic to P. interpunctella adults (LC₅₀ 0.99 mg/L air), while camphor, verbenol, borneol, and the two complete EOs exhibited lower toxicity. None of the EOs or main components exhibited significant toxicity against the larvae of P. interpunctella. This study provides evidence of the individual active substances accounting for the insecticidal activity of EOs from A. potaninii and A. fruticulosa. These EOs have potential as biological insecticides for controlling insect pest damage in stored crops.     

Characterization and PCR multiplexing of polymorphic microsatellite loci for the locust Locusta migratoria

Because of the scarcity of polymorphic genetic markers available in locust species, only a few population genetics studies have been carried out on this taxon. We isolated and characterized 11 polymorphic microsatellite loci in the pest locust Locusta migratoria capito, and described experimental conditions for polymerase chain reaction (PCR) multiplexing and simultaneously genotyping these loci. The number of alleles per locus ranged from six to 25, and the expected heterozygosity ranged from 0.431 to 0.957. Results of cross‐taxon amplification tests are reported in six other Locusta migratoria subspecies, six species of the Oedipodinae subfamily and two other pest locust species.     

Microsatellite markers for two species of dampwood termites in the genus Zootermopsis (Isoptera: Termopsidae)

Dampwood termites in the genus Zootermopsis inhabit forested areas in western North America. To better understand the colony composition and breeding structure of Zootermopsis, we identified polymorphic microsatellite loci to use in population analysis. Microsatellite loci were isolated from Zootermopsis nevadensis nevadensis (Hagen); however, all primers amplified homologous loci in Zootermopsis angusticollis (Hagen) and Zootermopsis nevadensis nuttingi (Hagen). Twelve loci were polymorphic in one or more of the above subspecies and species. The number of alleles per locus ranged from one to six, with some allelic differences among subspecies and species. We are currently utilizing the microsatellite markers to investigate the population genetics of Zootermopsis.     

Polymorphic microsatellite DNA markers from the Oriental Fruit Fly Bactrocera dorsalis (Hendel)

Six polymorphic microsatellite loci are isolated from the Oriental fruit fly Bactrocera dorsalis (Hendel), an agricultural pest in Asia, including Taiwan. To assess their potential utility as high‐resolution genetic markers, polymerase chain reaction (PCR) primers, amplification conditions, and an automated fluorescence detection protocol were developed. In analyses of 71 individual flies from six different areas of Taiwan, allele numbers ranged from five to 25 were detected for each locus. The observed heterozygosity ranged between 0.268 and 0.737 among these loci. No linkage disequilibrium was found. These microsatellite markers have potential utility to population structure and gene flow studies of B. dorsalis (Hendel).     

Di‐ and tri‐nucleotide repeat microsatellites for the mealy plum aphid,Hyalopterus pruni

Hyalopterus pruni is an invasive aphid pest in California. To study the population biology of this pest both in California and its native Mediterranean region, we have developed 11 di‐ and tri‐nucleotide repeat microsatellite markers. Each locus amplified in individuals representing the full range of geographical regions and host plants where Hyalopterus is found. Polymorphism was high, ranging from six to 22 alleles per locus in the individuals screened. These loci represent the first microsatellites developed for Hyalopterus and they should be of great value in studying the invasion biology and population structure of this insect pest.     

Isolation of additional polymorphic tri‐ and tetranucleotide microsatellite loci for the giant Australian cuttlefish (Sepia apama)

We isolated 10 polymorphic microsatellite loci for the giant Australian cuttlefish, Sepia apama, from a genomic library enriched for (AAC)_n and (AAAG)_n repetitive elements. In the nine loci that reliably amplified, the number of alleles ranged from four to 12 per locus with observed heterozygosity ranging from 0.343 to 0.926. These and a previously developed set of six loci will be useful for analysis of genetic structure of populations and determining input to a massive seasonal breeding aggregation in northern Spencer Gulf, Australia.     

Gene diversity at allozyme loci in the cottonwood leaf beetle,Chrysomela scripta

Gene diversity was studied in beetles collected from a poplar plantation at Ames, Iowa. The beetles are widely distributed throughout North America wherever poplars (Populus spp) occur. Of 38 loci interpretable by simple Mendelian criteria, 22 were polymorphic (58%). Nei's mean heterozygosity per locus was 20.1±4.0%. A mean 2.29±1.35 alleles per locus was detected. The foregoing levels of diversity are typical of Coleoptera. There were significant deviations from Hardy-Weinberg expectations at six loci, and of these loci, four deviations were judged to have been caused by technical problems in resolving heterozygotes. Twelve polymorphic loci are considered suitable to carry out studies on the breeding structure of this important pest.     

Isolation and characterization of the first microsatellite loci from the order Psocoptera in the economically important pest insect Liposcelis decolor (Pearman) and cross‐species amplification

A total of 11 microsatellite loci from the invasive insect pest Liposcelis decolor were isolated and characterized of which six loci were polymorphic. A population survey involving a total of 30–192 individuals per locus from five populations revealed a range of four to seven alleles per locus and moderate observed heterozygosities (0.183–0.565), highlighting the utility of these loci in further population genetic studies. Cross‐species amplifications were successful for two to 11 loci in five other Liposcelis species also of international economic importance.     

Microsatellite loci for tucumã of Amazonas (Astrocaryum aculeatum) and amplification in other Arecaceae

• Premise of the study: Microsatellite loci were developed for tucumã of Amazonas (Astrocaryum aculeatum), and cross-species amplification was performed in six other Arecaceae, to investigate genetic diversity and population structure and to provide support for natural populations management. • Methods and Results: Fourteen microsatellite loci were isolated from a microsatellite-enriched genomic library and used to characterize two wild populations of tucumã of Amazonas (Manaus and Manicoré cities). The investigated loci displayed high polymorphism for both A. aculeatum populations, with a mean observed heterozygosity of 0.498. Amplification rates ranging from 50% to 93% were found for four Astrocaryum species and two additional species of Arecaceae. • Conclusions: The information derived from the microsatellite markers developed here provides significant gains in conserved allelic richness and supports the implementation of several molecular breeding strategies for the Amazonian tucumã.     

准噶尔雅罗鱼组织同工酶表达特性

使用不连续PAGE法,分析了10尾准噶尔雅罗鱼(Leuciscus merzbacheri)眼睛、鳃、皮、背部肌肉、鳍和肝胰脏6种组织的10种同工酶(LDH,CCO,EST,CAT,POD,ME,MDH,G6PD,GDH,ADH)的差异表达,并对部分同工酶基因位点及表达酶谱表型进行了分析,以期为其种质资源保护和开发以及遗传育种等方面的研究提供基础资料。结果显示,10种同工酶中9种在6种组织中出现了明显的组织差异性,仅CCO在6种组织中的差异性较小。在对准噶尔雅罗鱼的10种同工酶的遗传多样性分析中,共记录到了21个基因位点,其中Est-1、Me-B、s-MDH、G6pd-A、G6pd-B和Adh-A为多态性基因位点。多态位点比例为:P=6/21=28.57%。     

Microsatellite analysis revealed a different approach of control of olive fly population (Bactrocera oleae) in Slovenia

The olive fruit fly (Bactrocera oleae Gmelin) is the most important olive pest in the north‐eastern Adriatic coast region. Despite the importance of olive production in the region, and the significance of the olive fruit fly, no information with respect to genetic diversity, population structure or dispersion patterns of this pest is available. The aims of this study were to investigate the genetic structure of the olive fly population in the Slovenian Istria region using microsatellite markers to determine olive fruit fly migration between locations and to establish an appropriate and effective strategy for controlling the pest population. Analysis was performed on a sample of 117 flies, collected from attacked olive fruits at three different locations. Olive fruit flies were genotyped using eight microsatellite loci. Sixty‐six alleles were identified over all microsatellite loci with an average of 8.25 alleles per locus. The population structure was determined with methods based on Bayesian principles using the BAPS 6.0 and STRUCTURE 2.3 programs. Genetic analysis confirmed unlimited migration and random mating between individuals of different microlocations, which suggests time‐coordinated first treatment in the region would be the best solution.     

Genetic rescue of an inbred captive population of the critically endangered Puerto Rican crested toad (<Emphasis Type="Italic">Peltophryne lemur</Emphasis>) by mixing lineages

The Puerto Rican crested toad (Peltophryne lemur) is currently composed of a single wild population on the south coast of Puerto Rico and two captive populations founded by animals from the northern and southern coasts. The main factors contributing to its decline are habitat loss, inundation of breeding ponds during storms, and impacts of invasive species. Recovery efforts have been extensive, involving captive breeding and reintroductions, habitat restoration, construction of breeding ponds, and public education. To guide future conservation efforts, genetic variation and differentiation were assessed for the two captive colonies and the remaining wild population using the mitochondrial control region and six novel microsatellite loci. Only two moderately divergent mitochondrial haplotypes were found, with one fixed in each of the southern and northern lineages. Moderate genetic variation exists for microsatellite loci in all three groups. The captive southern population has not diverged substantially from the wild population at microsatellite loci (F _ST = 0.03), whereas there is little allelic overlap between the northern and southern lineages at five of six loci (F _ST > 0.3). Despite this differentiation, they are no more divergent than many populations of other amphibian species. As the northern breeding colony may not remain viable due to its small size and inbred nature, it is recommended that a third breeding colony be established in which northern and southern individuals are combined. This will preserve any northern adaptive traits that may exist, and provide animals for release in the event that the pure northern lineage becomes extirpated.     

Breeding structure of an isolated cactophilic Drosophila population on a sandstone table hill

The effect of host cacti on the breeding structure of an isolated population of cactophilic Drosophila gouveai was studied. A comparison was made, using F statistics, of the allozyme frequencies at 11 loci among temporal samples of the adult population and six samples of progeny obtained from individual rotting cactus cladodes. The population appears to be structured by the cacti, forming breeding groups, and approximately four individuals contribute gametes to the progeny of each cladode. This D. gouveai population had a low degree of heterozygosity, compared with mean values for most Drosophila species, however it was within the range of values reported for other cactophilic Drosophila.     

Isolation and characterization of nuclear microsatellite loci in the anadromous marine fish Morone saxatilis

Molecular genetic studies of population structure and gene flow are commonly employed in fish stock assessment and breeding population delineation. Genetically structured breeding populations may have different effective population sizes, distinct reproductive rates and variable susceptibility to harvesting or breeding habitat degradation. Nine microsatellite loci were isolated for Morone saxatilis (Moronidae), an anadromous fish inhabiting the mid‐Atlantic. Microsatellite loci were isolated with a subtractive hybridization method and will be used to estimate population structure. The loci averaged 8.5 alleles each. Seven loci in the Choptank population and two loci in the Potomac population deviated from Hardy–Weinberg expected frequencies of heterozygotes.     

Development and characterization of polymorphic microsatellite markers in Linum usitatissimum

Thirty-five microsatellite loci were isolated and characterized in Linum usitatissimum using enriched genomic libraries. These loci were screened in eight cultivars from different countries and regions and were found to be polymorphic, with the number of alleles per locus ranging from two to six, and observed and expected heterozygosities ranging from 0.125 to 0.375 (mean 0.013) and from 0.233 to 0.842 (mean 0.601), respectively. These polymorphic new microsatellite loci will be useful for genetic linkage map construction, germplasm classification and identification, gene identification and quantitative trait loci mapping, and marker-assisted selection in breeding in L. usitatissimum.     

Isolation and characterisation of polymorphic microsatellite loci in the vulnerable spectacled flying fox, Pteropus conspicillatus

The spectacled flying fox, Pteropus conspicillatus, is listed as vulnerable in Australia and is under threat from numerous impacts. Primers to amplify eight co-dominant microsatellite loci were designed for Pteropus conspicillatus, based on an enriched genomic library. Four loci were monomorphic in this species while the remaining four loci were highly polymorphic with 16–23 alleles. Two of the four monomorphic loci were found to be polymorphic in Pteropus alecto, a closely related congener. All but one of the six polymorphic loci were in Hardy Weinberg equilibrium. Additionally, six microsatellite loci isolated for Pteropus rodricensis were tested against individuals of P. conspicillatus with all loci amplifying reliably. These loci will be used to investigate population genetic structure in the vulnerable spectacled flying fox.     

Dissection of genetic architecture for glucosinolate accumulations in leaves and seeds of Brassica napus by genome‐wide association study

Glucosinolates (GSLs), whose degradation products have been shown to be increasingly important for human health and plant defence, compose important secondary metabolites found in the order Brassicales. It is highly desired to enhance pest and disease resistance by increasing the leaf GSL content while keeping the content low in seeds of Brassica napus, one of the most important oil crops worldwide. Little is known about the regulation of GSL accumulation in the leaves. We quantified the levels of 9 different GSLs and 15 related traits in the leaves of 366 accessions and found that the seed and leaf GSL content were highly correlated (r = 0.79). A total of 78 loci were associated with GSL traits, and five common and eleven tissue‐specific associated loci were related to total leaf and seed GSL content. Thirty‐six candidate genes were inferred to be involved in GSL biosynthesis. The candidate gene BnaA03g40190D (BnaA3.MYB28) was validated by DNA polymorphisms and gene expression analysis. This gene was responsible for high leaf/low seed GSL content and could explain 30.62% of the total leaf GSL variation in the low seed GSL panel and was not fixed during double‐low rapeseed breeding. Our results provide new insights into the genetic basis of GSL variation in leaves and seeds and may facilitate the metabolic engineering of GSLs and the breeding of high leaf/low seed GSL content in B. napus.     

Isolation and characterization of microsatellite markers for Brachiaria brizantha (Hochst. ex A. Rich.) Stap

The first set of nuclear simple sequence repeat (SSR) loci for Brachiaria brizantha (Hochst. ex A. Rich.) Stap is described. A microsatellite-enriched library was constructed and 19 loci were characterized. About 13 SSR loci were found to be polymorphic and across-taxa amplification tests showed that six of them can be transferred to four other species of Brachiaria. This new SSR resource will be a powerful tool for population genetic studies of B. brizantha, for interspecific genetic studies within the genus Brachiaria, for mapping and for marker assisted selection in breeding.     

Competition and overlap of Oryzaephilus surinamensis and Plodia interpunctella populations under condition of stored date fruits

Plodia interpunctella and Oryzaephilus surinamensis are found in food storehouses including dates and palm storages. The current study aimed to determine competition and overlap potentials of the two pests of date fruits. Time series models were used to study two species populations and logistic growth model to estimate the effect of density of the species. The results revealed the environmental capacities of O. surinamensis and P. interpunctella were 433 and 1610 (maximum number per 20 g), respectively, and the population growth rates (r) were 1.2 and 1.3, respectively. Ecological balances of the two species were close to each other from the first to the third week. The population of O. surinamensis decreased in the fourth week of the competition. The highest population balance of the two species was in the 14th week. The potential of exploitable ecological niches (e_ij) and the amount of non-exploited ecological niches by any species (z_ij) for O. surinamensis was higher than for P. interpunctella from the 8th week untill the end of sampling period. The overlap of ecological niches in the two species (D) ranged from 0.94 to 1, indicating a complete overlap of temporal activity in the two populations on date palm. The current results of this study can be used by integrated pest management specialists. Information over the effects of species competition on population dynamics and their coexistence can be used to predict population status and to adopt simple pest control methods.